3-acyloxy-5,7,9(11)-pregnatrien-20-one-21-acetate adducts



Patented Dec. 9, 1952 UNITED STATES star the

S-ACYLOXY- 5,7,9 (11) -PRE GNATRIEN-ZO ONE-Zl-ACETATE ADDUCTS corporation of Michigan No Drawing. Application August 22, 1950, Serial No. 180,900

7 Claims.

C=O CH3 I wherein Ac is the residue of an organic carboxylic acid, especially such acids containing from one to eight carbon atoms, inclusive; and A is an adduct radical derived from a dienophile selected. from the group consisting of maleic acid, maleic anhydride, and maleic acid cliesters containing from one to eight carbon atoms, inclusive, in the esterifying group.

AcO-

It is an object of the present invention to provide a novel group of compounds which are useful in the preparation of steroid compounds containing an oxygen atom at carbon atom eleven. Another object of the invention is the provision of a process for the production of the novel compounds, adducts of 3-acyloxy-5,'7,9(11)-pregnatrien-20-one-2l-acetates. Other objects of the invention will become apparent hereinafter.

The compounds of the present invention, as previously stated, are useful in the preparation of steroid compounds having an oxygen atom attached to carbon atom eleven. Such compounds are of particular interest in the field of steroid research due to the biological activity of the cortical hormones and certain known derivatives thereof, which oxygenated steroids are known to have biological efiects differing markedly from the unoxygenated steroids. It is, therefore, of importance to investigate the oxygenated deriva tives of such adducts, particularly those oxygenated at carbon atom eleven, as well as to investigate the biological activity of the adducts themselves and their transformation products. The importance of such investigation is moreover emphasized by the acute shortage of adrenal cortiscal hormones, and the absence of any present suggestion for alleviation of the said shortage except through organic synthesis.

Novel compounds of the present invention which are of particular interest are those compounds of the above generic formula wherein Ac represents the residue of a carboxylic acid containing up to and including eight carbon atoms. Among the acids which can be used are formic, acetic, propionic, butyric, valeric, hexanois, heptanoic, octanoic, succinic, gutaric, cyclopentanoic, benzoic, toluic, and the like. Preferred acids are the lower-aliphatic acids. The acids may also contain substituents, such as halo, alkyl, and methoxy, which are non-reactive under the reaction conditions employed. The adduct bridge (A-) in such compounds may be represented by the graphic formula:

c-coon r o-( no which is representative of the maleic anhydride adduct.

The compounds of the invention are usually colorless crystalline. solids. The acidanhydride adducts are readily converted first to the 21- hydroxy acid and then to 3,21-dihydroxy acid adduct by hydrolysis. The acid adducts in turn are readily converted into diester adducts by esterification with conventional reagents such as the diazoalkanes [Wilds et al., J. Org. Chem. 13, 763 (1948)]. Conversely, the diesters may be hydrolyzed to the hydroxy dibasic acids which in turn may be converted into their corresponding anhydrides either by heat alone or preferably with a mild dehydrating agent such as acetic anhydride, in which case acylation of 3-hydroxy groups also occurs.

The compounds of the present invention have the formula:

CH C-CH;

C=0 CH:

theoretical, from one to four moles of lead tetrabe acetate per mole of adduct being preferred, and an organic solvent for the reaction is employed, for example acetic acid and water, benzene, or ether, with acetic acid being preferred. The reaction may conveniently be carried out at a tem perature between about twenty and 100 degrees centigrade, with temperatues of about -60 degrees centigrade, which are readily obtainable by employment of a water-bath, being preferred. The lead tetraacetate usually goes into solution quite readily, and the reaction is complete in from 24 to 96 hours. The 21-acetoxy adduct may then be recovered by evaporating the solution under a vacuum, dissolving the residue in ether, washing with Water, drying, and recrystallizing from a solvent, such as alcohol, if desired, or in other conventional manner.

The starting adducts of 3-acyloxy-5,7,9(1l)- pregnatrien-20-ones are conveniently prepared by the selective oxidation of enol ester of an adduct of 3-acyloxybisnor-5,7 ,9 l1) -cholatrien-22- al, represented by the formula:

wherein A and Ac have the values previously given.

Adducts of 3,22 diacyloxybisnor 5,7,9(11), 20(22)cholatetraenes [22-enol esters of B-acyloxybisnor-5,7,9(11) -cholatrien-22-als] are. conveniently prepared by subjecting an a'clduct of a 3 acyloxybisnor 5,7,9(11) cholatrien 22 al, of the formula:

Clla

,, wherein A and Ac have the values previously given, to the action of an acid anhydride or an acid halide in the presence of an alkaline salt of the acid. The starting adducts of 3-acyloxybisnor- 5,7,9(11) -cholatrien-22-als can be prepared from 20 adducts of 3-esters of dehydroergosterol by selective oxidation as described and claimed in the copending application Serial 111,100 of Robert H.

Levin, filed August 18, 1949, and as more fully described hereinafter.

The 3-esters of dehydroergosterol, from which the 3 acyloxybisnor 5,7,9(11) -cholatrien-22-al adducts are prepared, can be synthesized in several ways starting with ergosterol. For example, ergosterol can be transformed to dehydroergosterol with mercuric acetate according to known methods [Windaus et al., Ann. 465, 157 (1923) 'I and the 3-hydroxy group of the dehydroergosterol acylated by known procedure. Alternatively the B-hydroxy group of ergosterol can be acylatecl prior to the preparation of the dehydro derivative, a procedure which is particularly preferred in the preparation of the 3-acetoxy derivative. The adducts of dehydroergosterol are then prepared by the addition of maleic anhydride or the like to dehydroergosterol or a S-ester thereof according to known methods lHonigmann, Ann. 508, 89 (1934)]. The anhydrides can then be converted to their corresponding acids and esters if desired.

The ester group, when present in the 3-position of dehydroergosterol, is for the purpose of protecting the S-hydroxy group in subsequent chemical reactions. For this purpose any convenient ester of an organic carboxylic acid, which is nonreactive under the conditions of the reaction, is suitable. The preferred acids are the fatty acids such as formic, acetic, propionic, butyric, valeric. hexanoic, heptanoic, octanoic; dibasic acids such as malonic, succinic, phthalic; cycloaliphatic acids such as cyclopentanoic and cyclohexanoic; and aromatic acids such as benzoic, toluic, and the like. The acids may also contain substituents such as halogen, alkyl, the methoxy radical, and the like, and these substituents will be carried throughout the synthesis. If desired, the acyl group can be changed to another acyl group by saponifying the ester to give a 3-hydroxy compound, which can then be re-esterified as previously described.

A preferred method for preparing some of the dehydroergosteryl. adducts comprises the saponification of a 3-acyloxy-adduct of dehydroergosterol with dilute alkali followed by acidification. The 3-hydroxy dicarboxylic acid thus formed can be converted to the B-hydroxy anhydride by heat, or it can be converted to any desired 3acyloxy anhydride adduct by heating under reflux with the appropriate acid anhydride or chloride in pyridine solution. Dialkyl esters of the previously mentioned dicarboxylic acid adactive groups, such as halo,

ducts can be prepared by subjecting the acid to the action of an esterification reagent such as a diazoalkane [Wilds et al., J. Org. Chem. 13, 763 (1948) l, e. g., diazomethane, diazoethane, diazobutane, and the like.

The selective oxidation of an adduct of dehydroergosterol, or a 3-ester thereof, to produce an adduct of 3-hydroxybisnor-5,'7,9(11) -chloratrien- 22-al, or a 3-ester thereof, is accomplished by dissolving the dehydroergosteryl adduct in a suitable solvent, cooling to about minus 80 to plus degrees centigrade, and passing ozone into the solution until about 1.0 to 1.25 moles of ozone per mole of adduct have been absorbed. The temperature of the solution should be maintained below plus 30 degrees centigrade, preferably between a temperature of minus 30 and minus 70 degrees centigrade, during the addition of ozone, although temperatures as low as minus 80 and as high as plus 30 degrees centigrade are operative. The lower temperatures of the preferred range are readily obtained by cooling the solution of the adduct with a bath of solid carbon dioxide in acetone or the like, although various other methods of cooling can be used. Many of the customary solvents used in ozonizations such as chloroform, acetic acid, carbon tetrachloride, ethylene chloride, methylene chloride, and the like, can be used.

The ozonides are then decomposed under reducing conditions, that is, in the absence of oxidizing agents, whether added or formed in the course of the reaction by products of decomposition of the ozonide. This means that excess oxygen formed by decomposition of the ozonide is prevented from forming hydrogen peroxide by combining with any moisture present, and that molecular oxygen is prevented from oxidizing the aldehyde thus formed. This canbe conveniently accomplished by decomposing the ozonide in glacial acetic acid by the addition of finely-powclered zinc.

As is conventional with ozonizations when conducted in solvents, other than glacial acetic acid, the solvent used for ozonization is replaced, after completion of the ozonization, by adding glacial acetic acid and removing the lower-boiling solvent by fractional distillation. Alternatively, the solvent can be removed by careful warming under reduced pressure prior to the addition of glacial acetic acid, if desired.

After decomposition of the ozonide and removal of the zinc, the aldehyde can be recovered by diluting the acetic acid with water, or in other conventional manner, such as by formation of an aldehyde derivative, e. g., the dinitrophenylhydrazone.

Adducts of 3,22-diacycloxybisnor-5,l,9(l1) ,20- (22) -cholatetraenes [22-enol-esters of adducts of 3-acyloxybisnor-5,'7,9 (11) -cholatrien-22-alsl can be conveniently prepared by heating the corresponding 3-hydroxy or acyloxy aldehyde maleic acid anhydride, or maleic acid ester adduct with a large excess of an organic carboxylic acid anhydride in the presence of a small amount of the alkali metal salt of the acid corresponding to the anhydride employed or an acid catalyst such as para-toluene sulfonic or sulfuric acid. The preferred anhydride is acetic anhydride, but other anhydrides, such as propionic, butyric, valeric, hexanoic, and octanoic anhydrides, as well as benzoic acid anhydride, ortho-toluic acid anhydride, and the like, are also operative. The acid anhydrides can also be substituted by non-realkyl, and methoxy,

as in the case of chloroacetic. orthotoluic, or methoxybenzoic acid anhydrides. The reaction can be conveniently followed by observing the color changes in the reaction mixture, optimum yields being obtained by discontinuing the application of heat when the color of the solution changes from yellow to brown. Ordinarily the reaction is heated at about degrees centigrade for from about four to six hours, but temperatures as low as 100 and as high as degrees centigrade are also operative. The reaction is usually conducted at the boiling point of the anhydride, but in the case of the higher-boiling anhydrides, such as benzoic anhydride, a suitable temperature control, such as 100-150 degrees centigrade, must be used, since the adduct otherwise tends to decompose in the higher temperature range. If a S-hydroxy aldehyde adduct is thus reacted with an anhydride, the hydroxy group will be acylated, and, similarly, if a maleic acid adduct is used instead of a diester or an anhydride, the anhydride will be formed. The enol ester can be isolated by removing the excess anhydride under reduced pressure and separating the enol ester from alkali metal salts, which procedure yields a product sufliciently pure for most purposes, but which can be further purified by recrystallization from acetone-water, acetonepentane, or like solvents. if desired.

The ozonization of the thus-prepared enol acylate to produce a 3-acyloxy-5,7,9(11)-pregnatrien-ZO-one adduct involves dissolving the enol ester in a suitable solvent, cooling to about minus eighty degrees centigrade to plus thirty degrees centigrade, and passing ozone, ozonized air, or ozonized oxygen into the solution until about 1.0 to about 1.25 moles, preferably 1.0 to 1.1 moles, of ozone per mole of adduct have been absorbed. The addition of ozone to the 20:22 double bond is so rapid that only a small amount of ozone escapes from the reaction mixture and the amount of ozone ordinarily required therefore closely approximates the theoretical amount. Loss to the solvent, if any loss occurs, must be taken into consideration in calculating the amount of ozone to be introduced. The temperature of the solution should be maintained below plus thirty degrees centigrade, preferably between a temperature of minus thirty and minus seventy degrees centigrade, during the addition of ozone, although temperatures as low as minus eighty and as high as plus thirty degrees centigrade are operative. The lower temperatures of the range are readily obtained by cooling the solution of the adduct with a bath of solid carbon dioxide in acetone or the like, although various other methods of cooling may be employed. Many of the customary solvents used in ozonizations, such as chloroform, methylene chloride, ethylene chloride, carbon tetrachloride, acetic acid, and the like, can be used for the ozonization reaction.

The 20:22 ozonides thus produced are then decomposed under conditions normally employed for decomposition of such compounds. This can conveniently be accomplished by decomposing the ozonide with hydrogen peroxide, by hydrolysis, with zinc in glacial acetic acid, or by a catalytic amount of colloidal metal such as silver, platinum, or palladium in a solvent, such as glacial acetic acid, alcohol, or ethyl acetate, in which latter case reductive conditions, e. g., a hydrogen atmosphere, are also employed. The use of reductive conditions is well established in the art [Hill and Kelly, Organic Chemistry,

page 53, The Blakiston Company, Philadelphia (1934); Church et al., J. Am. Chem. Soc. 56, 176-184 (1934); Gilman Organic Chemistry, Second Edition, page 636, John Wiley and Sons, New York (1943); Long, Chem. Reviews 27, 452- 4-54 (1940)].

As is conventional with decomposition of ozonides with zinc, when the ozonizations are conducted in solvents other than glacial acetic .acid, the solvent used for the ozonization is recedure for the recovery of ketones, such as by H formation of a carbonyl derivative, e g., the 2,4- dinitrophenylhydrazone. Recrystallization from acetone or the like results in a more highly purified'ketone product.

The following examples are illustrative of the process and products of the present invention, but are not to be construed as limiting.

PREPARATION l.DIMETI-IYL MALEATE ADDUC'r or DEHYnnoERcosTisnYL BENZOATE To a solution of 21 grams of dimethyl maleate adduct of dehydroergosterol in 69 milliliters of warm pyridine was added 9.5 milliliters of benzoyl chloride. After standing at room temperature for fifteen minutes, the mixture was poured into 1400 milliliters of ice-water and the solid removed by filtration, dried, and recrystallized from acetone. There was thus obtained 26.4 grams of dimethyl maleate adduct of dehydroergosteryl benzoate, melting at 203 to 205.5 degrees centigrade.

Pmammrr N 2.-DIMETHYL laLiLEATn ADDUCT or DEnYDnoERsosTERYL ACETATE In a manner essentially that described in Preparation 1, the dimethyl maleate adduct of dehydroergosteryl acetate, melting at 177 to 1'79 degrees centigrade, was prepared from th dimethyl maleate adduct of dehydroergosterol and acetyl chloride.

PREP. .RATION 3.DIMETHYL MALEATE ADDUCT or DEHYDROERGOSTERYL FORMATE A solution of six grams of dimethyl maleate adduct of dehydroergosterol in fifty milliliters of 87 percent formic acid was heated under reflux for one hour, cooled, and the dimethyl maleate adduct of dehydroergosteryl formate filtered therefrom. Upon crystallization from acetone, the purified material melted at 177.5 to 178.5 degrees centigrade.

PREPARATION 4.MALEI Acn) Anpuo'r or DEHYDROERGOSTEROL A solution of 2.0 grams of sodium hydroxide in twenty milliliters of water was'added to a solution of 1.73 grams of the maleic anhydride adduct of dehydroergosteryl acetate (M. P. 230- 232 degrees Centigrade) in forty milliliters of dioxane. The mixture solidified, but dissolved on addition of 300 milliliters of water and heating to eighty degrees centigrade. After half an hour the solution was cooled and made acid with aqueous three normal hydrochloric acid, to give 1.61 grams of precipitate. On crystallization from a dioxane-water mixture, the maleic 8 acid adductof dehydroergosterol melted at 192 degrees centigrade.

PREPARATlON 5..\Lu.r1c Axnromns Author or 3-HEPTANoYLoXYnEHrnnoEnoosrERoL The maleic acid adduct of dehydroergosterol from Preparation 4 was dissolved in a mixture of seven milliliters of warm pyridine and fourteen milliliters of heptylic anhydride, and the mixture heated under reflux for one hour. About eighty percent of the reaction solvent was removed under reduced pressure, and the residue then dissolved in methyl alcohol. The methyl alcohol solution was concentrated and cooled to yield 4.8 grams of the maleic anhydride adduct of 3 heptanoyloxydehydroergosterol, melting at 186-1915 degrees centigrade.

PREPARATION G.\LiLEro Azvnrmuma Annocrr or 3- BETA-ACETOXYBISNOR-5,7,9 (11) -CHOLATltlEN-22-AL solution of 5.35 grams of the maleic anhydride adduct'of 3-beta-acetoxydehydroergosterol in 107 milliliters of methylene chloride was cooled to about minus seventy degrees centigrade and ozonized until 505 milligrams of ozone had been absorbed. The temperature of the solution was then gradually raised to about plus ten to fifteen degrees centigrade, whereupon seventy milliliters of glacial acetic acid was added and the methylene chloride removed under reduced pressure. Seven grams of zinc dust Was then added to the cold solution at a uniform rate over a period of ten minutes, while keeping the reaction temperature below. plus twenty degrees Centigrade. After being stirred for fifteen minutes, the mixture wasfiltered and the filtrate poured into water. There was thus obtained 4.31 grams of maleic anhydride adduct of 3-betaacetoxybisnor 5,7,9(11) cholatrien 22 a1, a fine white powder which melted at 187-197 degrees centigrade.

To a solution of 0.30 gram of the maleic anhydride adduct of 3-beta-acetoxybisnor-S,7,9- (11)-cholatrien-22-al, in thirty milliliters of ethanol, was addedtwenty milliliters of alcohol containing one percent 2,4 dinitrophenylhydrazine and three percent concentrated hydrochloric acid. The mixture was allowed to stand for one hour at room temperature and then placed in a refrigerator to complete precipitation of the yellow crystals. The precipitate was then collected and recrystallized from a mixture of chloroform and alcohol, to give the ZA-dinitrophenylhydrazone of the maleic anhydride adduct of 3-beta-acetoxybisnor-5,7,9(11)-cholatrien-22- a melting a 269-271 degrees centigrade.

PREPARATION 7.l\IALEIC ANHYDHIDE Annual or 3- BETA-ACETOXYBISNGit-5,7,9 (11) -CHOLATRIEN-22-AL A two-liter, round-bottom flask was charged with fifty grams (0.93 mole) of dehydroergosteryl acetate maleic anhydride adduct and one liter of methylene chloride. The solution was cooled to Dry-Ice temperature with a tricholoroethylene bath and ozonized oxygen passed through at. a

'rate of 1200 milliliters of oxygen per minute (at this rate the ozonizer was producing about 36 over in vacuo at forty degrees centigrade or below. The flask was then placed in a water bath and fitted with a stirrer. An additional 200 milliliters of acetic acid was added and the ozonide decomposed by the addition of fifty grams of zinc dust. The zinc dust was added in portions over a period of twenty to thirty minutes while the solution was stirred and the temperature maintained at seventeen to twenty degrees centigrade. After addition, the mixture was stirred for another twenty minutes and then filtered. The precipitated zinc dust was washed by filtering 100 milliliters of acetic acid therethrough, and the filtrate gradually diluted with water (1100 to 1200 milliliters) until the product had been drowned out. The product was then cooled in the refrigerator overnight and filtered. The yield of crystalline product was 42 grams, assaying 89-95 percent of the desired aldehyde.

PREPARATION 8 In a manner essentially that described in Preparation 6, the following compounds were prepared.

(1) Maleic anhydride adduct of 3-beta-formoxybisnor-5,7,9 (11) -cholatrien-22-al, melting at 95-130 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 165-168 degrees centigrade.

(2) Maleic anhydride adduct of 3-beta-heptanoyloxybisnor 5,7,9(11) oholatrien 22 al, melting at 197.5-199 degrees centigrade. 2,4- dinitrophenylhydrazone, melting at 253-257 degrees centigrade.

(3) Dimethyl maleate adduct of B-beta-benzoyloxybisnor 5,7,9(11) cholatrien 22-al, melting at 183-187 degrees centigrade. 2,4- dinitrophenylhydrazone, melting at 224-249 degrees centigrade.

(4) Dimethyl maleate adduct of 3-beta-acetoxybisnor-5,'7,9(11)-cholatrien-22-al, melting at 172-178 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 238 to 244 degrees centirade.

(5) Dimethyl maleate adduct of 3-hydroxybisnor 5,7,!)(11) cholatrien 22 a1, melting at 163-170 degrees centigrade. 2,4-dinitrophenylhydrazone, melting at 250-254 degrees centigrade.

In a manner similar to the above, the maleic anhydride adduct of 3-hydroxybisnor-5,7,9(11)- cholatrien-ZZ-al is obtained from dehydroergosteryl maleic anhydride adduct; the maleic acid adduct of 3hydroxybisnor-5,7,9(11)-cholatrien-22-al is obtained from dehydroergosteryl maleic acid adduct; and 3acyloxybisnor-5,7,9- (11) -cholatrien-22-al maleic acid adducts are obtained from the maleic acid adduct of B-acyloxydehydroergosterols.

PREPARATION 9.-DIMETHYL MALEATE ADDUCT or 3-HYnRoXYBIsNoR-5,7,9 (11) -CHOLATRIEN22-AL cooling in a water-bath at fifteen degrees centigrade, four grams of zinc dust was added in por- 10 tions with stirring, the temperaure being maintained between fifteen and twenty degrees centigrade. Stirring was continued for another fifteen minutes, whereai-ter the zinc was separated by filtration. The filtrate was diluted with water to cloudiness, extracted with ether, the ether extract washed with sodium bicarbonate and then with water to neutrality, the solution then dried over sodium sulfate and evaporated to dryness in vacuo. acid and water, giving 1.92 grams (81.5 percent of the theoretical), melting point 91-97 degrees centigrade, which yielded a dinitrophenylhydrazone derivative in 72.5 percent yield, melting point 212-238 degrees centigrade. The aldehyde was recrystallized and found to have a purified melting point of 163-170 degrees centigrade, while the dinitrophenylhydrazone derivative was recrystallized until a melting point of 250-254 degrees centigrade was attained.

PREPARATION 10.-MALEI0 ANHYDRIDE ADDUCT 0F 3- BETA-ACETOXY-22ACETOXYBISN 0R-5 ,7 ,9 11) ,20 2-2 CHOLATETRAENE A mixture of twenty grams of the maleic anhydride adduct of S-beta-acetoxybisnor- 5,7,9(11)-cholatrien-22-al, six grams of anhydrous sodium acetate, and 600 milliliters of acetic anhydride, was heated under reflux for six hours, whereafter volatile components were removed under reduced pressure. The resulting solid was digested with five fifty-milliliter portions of boiling acetone for five minutes each, and the extracts combined and diluted with milliliters of water. There was thus obtained sixteen grams of the maleic anhydride adduct of 3-beta-acetoxy 22 acetoxybisnor 5,7,9(11) ,20(22) cholatetraene, which melted at 186 to 193 degrees centigrade. Recrystallization of the crude product from a mixture of acetone and pentane raised the melting point to 200.5 to 202 degrees centigrade.

PREPARATION 11 In a manner essentially that described in Preparation 10, the following compounds were prepared:

(1) The dimethyl maleate adduct of 3-betabenzoyloxy 22 acetoxybisnor-5,7,9(11) ,20(22) cholatetraene, which melted at 210 to 211 degrees Centigrade.

(2) The dimethyl maleate adduct of 3-betaacetoxy 22 acetoxybisnor 5,7,9(l1),20(22)- cholatetraene, which melted at 181 to 183 degrees centigrade.

In the same manner as given above, 22-acy1oxy, e. g., formoxy, acetoxy, propionoxy, butyroxy, valeroxy, hexanoyloxy, heptanoyloxy, octanoyloxy, 'benzoyloxy, naphthoyloxy, and the like 3- acyloxybisnor 5,7,9(11),20(22) cholotetraene adducts, are obtained from the compounds of Preparations 6, 7, and 8. Such representative compounds include 3-formoxy-22-acetoxybisnor- 5,7 ,9(11) ,20 (22) -cholatetraene, 3-propionoxy-22- acetoxybisnor 5,7,9(1l),20(22) cholatetraene, 3,22 dipropionoxybisnor 5,7,9(11),20(22) -cho latetraene, 3,22 dibenzoyloxybisnor 5,7,9(11) 20(22) -cho1atetraene, and 3-heptanoyloxy-22- octanoyloxy-bisnor-5,7,9 (11) ,20 (22) cholatetraene adducts with maleic anhydride or maleic acid esters such as the dimethyl maleate, diethyl maleate, dipropyl maleate, diisopropyl maleate, dibutyl maleate, dioctyl maleate, dibenzyl maleate, and the like.

The residue was crystallized from acetic A solution of 5.08 grams of the mal ic anhydride adduct of 3-beta-acetoxybisnor-5,7,9(11)- cholatrien-22-al enl acetate in 100 milliliters of methylene chloride was cooled to about minus seventy degrees centigrade and ozonized until 483 milligrams of ozone had been absorbed. Fifty milliliters of glacial acetic acid was then added and the methylene chloride removed under reduced pressure. An additional thirty milliliters of glacial acetic acid was then added and the ozonide decomposed by adding seven grams of powdered zinc at a substantially uniform rate while maintaining the reaction temperature between seventeen and twenty degrees centigrade. The mixture was stirred for an additional twenty minutes, filtered, and the zinc washed with 140 milliliters of glacial acetic acid. The organic extracts were combined and diluted with seventy milliliters of water. When crystallization commenced, the rate of precipitation was increased by addition of two volumes of water. There was thus obtained 4.0 grams of the maleic anhydride adduct of 3-beta-acetoxy-5,7,9(11)-pregnatrien- 20-one, which melted at 240 to 264.5 degrees centigrade. Several recrystallizations of the crude material from acetone raised the melting point to 263.5 to 264.5 degrees centigrade.

PREPARATION 13.MALEI ACID ADDUcT or 3-mon- HYDRoxY-5,7,9 (11)-PREGNATRIEN-20ONE A solution of 4.52 grams (0.0100 mole) of the maleic anhydride adduct of 3-beta-acetoxy- 5,7,9(ll)-pregnatrien--one, M. P. 263-2645 degrees centigrade, in a mixture of 100 milliliters of 1,4-dioxane and 400 milliliters of water containing four grams (0.10 mole) of sodium hydroxide was allowed to stand at room temperature for two and one-half hours, whereupon a small quantity of plate-like crystals formed. These were dissolved by heating the mixture to seventy degrees centigrade for one-half hour. The reaction mixture was then made acid with fifty milliliters of three normal hydrochloric acid and refrigerated to give a precipitate of 3.05 grams of needle-like crystals melting at 173-177 degrees centigrade. On crystallization from a dioxane-water mixture, the compound melted at 211-215 degrees centigrade. was found to vary somewhat with the rate of heating.

Analysis:

Calculated for CH32O6 C, 70.07; H, 7.53 Found C, 69.80; H, 7.47

PREPARATION 1l.DI.\I1:r1IYL MALIJATE or 3-min- HYoRo: :Y-5,7,9 (11)-PREGNATRlEN-20-ONE A suspension of 0.400 gram of the maleic acid adduct of 3-beta-hydroxy-5,7,9(11) -pregnatrien- 20-one, in fifty milliliters of dry ether, was cooled in an ice-salt bath while a slight excess of diazomethane in methylene chloride was added over a 25 minute period with stirring. Ten minutes after addition was complete, the solution was placed on a steam bath and concentrated rapidly to dryness. The residue was crystallized from an acetone-water mixture to give 0.34 gram of the dimethyl maleate of S-beta-hydroxy- 5,7,9(ll) -pregnatrien-20-one, melting at 193-195 degrees centigrade. After chromatography and recrystallization, the compound melted at 192- 197 degrees centigrade.

In the same manner as given above, other dialkyl maleates, e. a, the diethyl, dipropyl, diiso- The melting point a propyl, dibutyl, and dioctyl maleates of 3-hydroxy-5,7,9(ll) -pregnatrien-20-one are prepared from 3-hydroxy-5,7 ,9(11) -pregnatrien 20 one maleic acid adduct and the appropriate diazoalkane, or by other equivalent esterification procedure.

PREPARATION l5.DIMETlIYL MALEATE or 3-BlCTA- Acxcroxr-SJQ 11) -IRECNATRlEN-20-ONE centigrade.

Analysis:

Calculated for C29H3aO'1 C, 69.86; H, 7.68 Found C, 69.81; H, 7.86 69.70; 7.62

By the same manner of esterification, the following C-3 esters were prepared: (1) dimethyl maleate adduct of 3 beta formoxy-5,7,9(11) pregnatrien-ZO-one, melting point 223-230 degrees centigrade, and (2) the dimethyl maleate adduct of 3-beta-benzoyloxy-5,7,9(11)pregnatrien-ZO-one, melting point 250-254 degrees centigrade.

PREPARATION l0.MALEIc ANIIYDRIDE AnnUC'r or 3- BETAHEPTANOYLOXY-5,7 ,9 (11) -PREGNATRIEN-20-ONE lhe maleic anhydride adduct of 3-beta-heptanoyloxy-5,7 ,9 11) -pregnatrien-20-one, melting point -171 degrees centigrade, was prepared by refluxing the maleic acid adduct of 3-betahydroxy-5,7,9(ll) -pregnatrien-20-one with heptylic anhydride and pyridine for a period of twenty hours, and working up the reaction product in the usual manner.

PREPARATION 1T.-1\IALEIC ANHYDRIDE Annual or 3-BETA-I1YDROXY-5,7,9 (11) -PREGNATRIEN-20-ONE Similarly, the maleic anhydride adduct 0f 3- beta-hydroxy 5,7 ,9(11) pregnatrien 20 one, melting point about degrees centigrade, was prepared by refluxing the maleic acid adduct of 3 beta hydroxy 5,7,9 (11) -pregnatrien-20one with Dowtherm for eight hours. The 3-hydroxymaleic anhydride adduct is also obtained by heating the 3-hydroxy maleic acid adduct to just above its melting point, which procedure causes water to be evolved, with the closing of the anhydride ring.

In the same manner as given above, still other 5,7,9(ll) -pregnatrien-20-one adducts are prepared from the corresponding 3,22-diacyloxybisnor-5,7,9(11),20(22)-cholatetraene maleic acid, maleic acid anhydride, and maleic acid diester adducts. Such compounds include the 3-formoxy-5,7,9(l1) -pregnatrien 20 one maleic acid, maleic acid anhydride, dimethyl maleate, diethyl maleate, dibutyl maleate, dioctyl maleate, diisopropyl maleate, dibenzyl maleate, and like adducts; the corresponding 3-propi0noxy, butyroxy, valeroxy, hexanoyloxy, heptanoyloxy, octanoyloxy, naphthoyloxy, benzoyloxy, and similar 20-ketone adducts, including, for example, 3-propionoxy-5,7,9(11) -pregnatrien-20-one dipropyl maleate, 3 benzoyloxy-5,7,9(ll)-pregnatrien-20-one dibenzoyl maleate, 3 heptanoyloxy-5,7,9(l1)- 13 pregnatrien--one dimethyl maleate, 3-valeroyloxy-5,'7,9(11) -pregnatrien-20-one maleic acid anhydride adducts, and the like.

Example 1.-Maleic anhydride adduct of 3-beta-, 21 -diaceto:cy-5,7 ,9 (11 -pTegnatrien-20-one The maleic anhydride adduct of 3-beta-acetoxy-5,7,9(l1)-pregnatrien 20 one (0.45 gram, .001 mole) and 1.33 gram of lead tetraacetate were added to milliliters of 98.3 percent acetic acid which had been distilled over chromium trioxide. The mixture was heated in a water bath maintained at about 56 degrees centigrade and allowed to stand at that temperature for seventytwo hours. The lead tetraacetate dissolved slowly during the first hour and went into solution. The solution was then evaporated in vacuo until only a few milliliters of solvent remained, the residue was dissolved in ether, Washed with water, dried, and evaporated to dryness. Crystallization of the residue from alcohol gave 0.17 gram of maleic anhydride adduct of 3-beta-,21- diacetoxy-5,'l,9 (11) -pregnatrien-20-one, melting at 225 to 235 degrees centigrade. After several recrystallizations from methanol, the material melted at 240-243 degrees centigrade.

Analysis:

Calculated for Casi-13403"..- C, 68.22; H, 6.71 Found C, 68.28; H,6.75

Example 2.-Dimethyl maleate adduct of 3- beta 21 diacetomy 5,7,9(11) pregnatrien-ZO-one In the manner of the preceding example, the dimethyl maleate adduct of S-beta-acetoxy- 5,7,9(11) -pregnatrien-20-one was converted to the dimethyl maleate adduct of 3-beta-,21-diacetoxy 5,7,9(11) pregnatrien 20 one, which melted at 210-214 degrees centigrade.

Analysis:

Calculated for C3lH4009 C, 66.89; H, 7.14 Found C, 66.72; H, 7.24: 67.09; 7.34

Example 3.-3 beta acetoxy 21 hydroxy- 5,7,9(11)-p'regnutrien-20-one maleic acid adduct One gram of 3-beta-,2l-diacetoxy-5,7,9(11)- pregnatrien-20-one maleic anhydride adduct was dissolved in 100 milliliters of dioxane and the solution cooled to just above the freezing point of dioxane. A second solution consisting of 25 milliliters of 1.5 normal sodium hydroxide solution and 350 milliliters of water was cooled in an ice-bath and then added to the dioxane solution. The volume of the reaction mixture was then brought up to 500 milliliters by addition of water and allowed to stand at zero degrees centigrade for one hour. The speed of the saponiflcation is followed by removing aliquots at different time intervals and titrating with standard hydrochloric acid. At one hour, three equivalents of base had been used up. The reaction mixture was made acid to Congo Red paper with ten percent hydrochloric acid and was concentrated n the steam bath in vacuo to about one hundred milliliters. This causes crystallization to take place, and, after cooling, 0.61 gram of 3 beta acetoxy 21, hydroxy- 517,9(11)pregnatrien-20-one maleic acid adduct, melting point 235-240 degrees centigrade, was obtained.

Example 4.-3 beta acetoxy 21 hydroxy- 5,7,9(11) -preynatrz'en-20-one maleic anhydride adduct Without further purification the acid thus produced in the preceding example was placed in a vacuum oven and heated for ten hours at 136-140 degrees centigrade at eight to ten millimeters of pressure. The resulting product was recrystallized from acetone-water and gave 0.43 gram, melting point 225-233 degrees centigrade, of 3 beta acetoxy 21 hydroxy 519(11)- pregnatrien-ZO-one maleic anhydride adduct. After two recrystallizations from ethyl acetatehexane, the product melted at 244-248 degrees centigrade. Infra red analysis conclusively showed that the 21-acetoxy group had been removed.

Example 5.-3 beta 21 dihydromy 5,7,9(11) pregnatrien-20mm malez'c acid and anhydride adducts If in the above procedure of Example 3, the saponification is allowed to go longer, until four equivalents of base are used, both the 3 and 21 acetoxy groups are removed and the 3beta-,21- dihydroxy-5,7,9 (11) -pregnatrien-20-one maleic acid adduct is produced, which can be converted to the anhydride by heating in a vacuum according to the manner of the preceding examp e.

In the same manner as given for all of the foregoing maleic anhydride adducts, the corresponding maleimide and N-alkylmaleimide adducts are prepared, starting from the selected 3 acyloxy-5,'7,9(11) pregnatrien 20 one maleimide or N-alkylmaleimide adduct.

It is to be understood that the invention is not to be limited to the exact details of operation or exact compounds shown and described, as obvious modifications and equivalents will be apparent to one skilled in the art, and the invention is therefore to be limited only by the scope of the appended claims.

We claim:

1. A 3 substituted 5,7,9(11)-pregnatrien- 20-one-21-substituted adduct of the formula:

CHa

CH2-OX lected from the group consisting of hydrogen and the residue of an unsubstituted monocarboxylic aliphatic acid containing from one to eight carbon atoms, inclusive. and wherein A is an adduct radical of a dienophile of the group consisting of maleic acid, maleic anhydride, and di-lower alkyl esters of maleic acid, wherein the esterifying groups contain from one to eight carbon atoms, inclusive.

2. .A 3 acyloxy 5,7,9(l1) pregnatrien- 20-one-21-acetate maleic anhydride adduct, wherein the acyloxy group is of the formula AcO, .Ac being the residue of an unsubstiuted monocarboxylic aliphatic acid containing from one to eight carbon atoms, inclusive.

3. 3,21 diacetoxy 5,7,9(11) pregnatrien- 20-one maleic anhydride adduct.

4. 3,21 diacetoxy 5,7,9(11) pregnatrien- 20-one dimethyl maleate adduct.

16 5. 3-acetoxy 21 hydroxy 5.7.901) pregnatrien-ZO-one maleic anhydride adduct.

6. 3 acetoxy 21 hydroxy 5,7,9(11) pregnatrien-ZO-one maleic acid adduct.

7. 3,21 dihydroxy 5,7,9(11) pregnatrien- 20-0ne maleic anhydride adduct.

ROBERT H. LEVIN. A VERN McINTOSI-I, JR. GEORGE B. SPERO.

Name Date Bockmuhl Feb. 4, 1941 Number 

1. A 3 - SUBSTITUTED - 5,7,9(11)-PREGNATRIEN20-ONE-21-SUBSTITUTED ADDUCT OF THE FORMULA: 